In vitro culture and autoª²contraction of Spragueª²Dawley rats¡¯ skeletal myoblasts

    ZHAO Keª²Yan, YU Shiª²Qiang, CAI Zhenª²Jie, WU Tieª²Jun, HU Jun, ZHOU Heª²Ping

    Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi¡¯an 710033, China

    ¡¾Abstract¡¿ AIM: To establish a practical procedure for culturing Spragueª²Dawley rats¡¯ myoblasts in vitro and to describe the characteristics of these myoblasts. METHODS: Muscle samples were minced and digested by collagenase¢ñand purified with Percoll gradient liquid. Myoblasts were tested by ¦Áª²actin and desmin immunochemistry stains. Many of the myoblasts could autoª²contract, so isoproterenol (IP) was added into the culture bottles with the final density as 0.1¦Ìmol/L so as to compare the contracting rates. RESULTS: The myoblast viability was measured as up to 90 % by trypan blue. Cell proliferation reached the peak 3-5 d after the cells were planted, and cell purity could be as high as 95 % by ¦Áª²actin and desmin immunochemistry stains tests. Myoblastes could contract on the 2nd or 3rd d after they were fused and the contracting rates ranged from 50 to 160 beats per minute for the most part, while some contracted with 200 beats per minute. When isoproterenol was added, the mean contracting rate increased about 18.4 beats per minute compared with that before the adding isoproterenol. CONCLUSION: Myoblasts can autoª²contract and IP can increase their contracting rates. The transplant of myoblasts into the ischemia heart has been proved to be effective in the treatment of heart diseases. ......